Professional Documents
Culture Documents
Senior Editors
Owen R. Fennema University of WisconsinMadison
Y.H. Hui Science Technology System
Marcus Karel Rutgers University (emeritus)
Pieter Walstra Wageningen University
John R. Whitaker University of CaliforniaDavis
Additives P. Michael Davidson University of TennesseeKnoxville
Dairy science James L. Steele University of WisconsinMadison
Flavor chemistry and sensory analysis John H. Thorngate III University
of CaliforniaDavis
Food engineering Daryl B. Lund University of WisconsinMadison
Rickey Y. Yada
University of Guelph
1. Flavor Research: Principles and Techniques, R. Teranishi, I. Hornstein, P. Issenberg, and E. L. Wick
2. Principles of Enzymology for the Food Sciences, John R. Whitaker
3. Low-Temperature Preservation of Foods and Living Matter, Owen R.
Fennema, William D. Powrie, and Elmer H. Marth
4. Principles of Food Science
Part I: Food Chemistry, edited by Owen R. Fennema
Part II: Physical Methods of Food Preservation, Marcus Karel, Owen
R. Fennema, and Daryl B. Lund
5. Food Emulsions, edited by Stig E. Friberg
6. Nutritional and Safety Aspects of Food Processing, edited by Steven
R. Tannenbaum
7. Flavor Research: Recent Advances, edited by R. Teranishi, Robert A.
Flath, and Hiroshi Sugisawa
8. Computer-Aided Techniques in Food Technology, edited by Israel
Saguy
37. Omega-3 Fatty Acids in Health and Disease, edited by Robert S. Lees
and Marcus Karel
38. Food Emulsions: Second Edition, Revised and Expanded, edited by
Kre Larsson and Stig E. Friberg
39. Seafood: Effects of Technology on Nutrition, George M. Pigott and
Barbee W. Tucker
40. Handbook of Vitamins: Second Edition, Revised and Expanded,
edited by Lawrence J. Machlin
41. Handbook of Cereal Science and Technology, Klaus J. Lorenz and
Karel Kulp
42. Food Processing Operations and Scale-Up, Kenneth J. Valentas,
Leon Levine, and J. Peter Clark
43. Fish Quality Control by Computer Vision, edited by L. F. Pau and R.
Olafsson
44. Volatile Compounds in Foods and Beverages, edited by Henk Maarse
45. Instrumental Methods for Quality Assurance in Foods, edited by
Daniel Y. C. Fung and Richard F. Matthews
46. Listeria, Listeriosis, and Food Safety, Elliot T. Ryser and Elmer H.
Marth
47. Acesulfame-K, edited by D. G. Mayer and F. H. Kemper
48. Alternative Sweeteners: Second Edition, Revised and Expanded, edited by Lyn O'Brien Nabors and Robert C. Gelardi
49. Food Extrusion Science and Technology, edited by Jozef L. Kokini,
Chi-Tang Ho, and Mukund V. Karwe
50. Surimi Technology, edited by Tyre C. Lanier and Chong M. Lee
51. Handbook of Food Engineering, edited by Dennis R. Heldman and
Daryl B. Lund
52. Food Analysis by HPLC, edited by Leo M. L. Nollet
53. Fatty Acids in Foods and Their Health Implications, edited by Ching
Kuang Chow
54. Clostridium botulinum: Ecology and Control in Foods, edited by
Andreas H. W. Hauschild and Karen L. Dodds
55. Cereals in Breadmaking: A Molecular Colloidal Approach,
Ann-Charlotte Eliasson and Kre Larsson
56. Low-Calorie Foods Handbook, edited by Aaron M. Altschul
57. Antimicrobials in Foods: Second Edition, Revised and Expanded,
edited by P. Michael Davidson and Alfred Larry Branen
58. Lactic Acid Bacteria, edited by Seppo Salminen and Atte von Wright
59. Rice Science and Technology, edited by Wayne E. Marshall and
James I. Wadsworth
60. Food Biosensor Analysis, edited by Gabriele Wagner and George G.
Guilbault
61. Principles of Enzymology for the Food Sciences: Second Edition, John
R. Whitaker
62. Carbohydrate Polyesters as Fat Substitutes, edited by Casimir C.
Akoh and Barry G. Swanson
63. Engineering Properties of Foods: Second Edition, Revised and
Expanded, edited by M. A. Rao and S. S. H. Rizvi
Preface
To establish the effect that any nutrient has on human physiology takes
years of research, punctuated by ongoing scientific debate. The task of determining what benefits can be derived from eating complex carbohydrates is no less
rigorous. The term complex carbohydrates became popular in February of
1977, when the U.S. Senate Committee on Human Needs and Nutrition loosely
indicated it as available starch. In their report, the committee encouraged liberal
consumption of complex carbohydrates to achieve better health. In 1991 the U.S.
Food and Drug Administration (FDA) proposed listing its quantities on food
labels (as starches and any dextrins of 10 or more saccharide units). However,
when the final regulations were issued in 1993, complex carbohydrates were not
included in the labeling guidelines. During the 2-year interim, the FDA and U.S.
Department of Agriculture (USDA) had concluded that although increased consumption of complex carbohydrates was desirable, neither a specific definition
of nor a methodology for analyzing complex carbohydrates was clearly enough
established to commence the labeling. The 1995 USDA dietary guidelines continued to recommend diets high in complex carbohydrates (defined as starch and
dietary fiber). Yet the tool that would ultimately inform consumers about the levels
of complex carbohydrates in their food (the nutrition label) still does not exist.
Because less conflict occurred in defining dietary fiber, an analytical
methodology was developed, validated, and has been now used in nutrition labeling since the 1980s. Labeling for complex carbohydrates, on the other hand, has
not yet achieved its place in consumer nutrition education.
As with any scientific issue that results in debate and public policy, each
idea and method about how to define and analyze complex carbohydrates has its
supporters, detractors, and those whose opinions vacillate (sometimes frequently). Current efforts to educate the public about the health benefits of complex carbohydrates through publicity and food labeling continue to be offset, in
part, by individuals and groups who want the term discontinued. Such arguments
are similar to ones heard in the course of creating a methodology and labeling
system for dietary fiber, although there was more universal agreement.
In 1990, after initially supporting the term dietary fiber, members of a
task force from the British Nutrition Foundation decided that they wanted instead
to make the term obsolete in scientific literature. They wanted to refer to just a
portion of dietary fiber, the entity non-starch polysaccharides. However, their
position was not accepted among international scientists and the term dietary
fiber prevailed. Today, it is accepted and used even by members of the task
force. Books and research continue to focus on dietary fiber and reinforce the
importance of a liberal dietary fiber intake. One need only look at the number
of workshops and symposia to realize that the term and its definition will continue
to form the base of discussions for years to come.
Mirroring this earlier debate over dietary fiber, a small but vocal number
of scientists want the term complex carbohydrates to be discontinued. These
scientists are suggesting that because of unresolved differences of opinion on
small details regarding the makeup of complex carbohydrates, we should throw
out the term and ignore the important strides that have been made in understanding this important component of our diet. However, the majority of scientists
working in the field support a continued effort in researching analytical methodologies and physiological effects even though the definition of complex carbohydrates remains loose. The term dietary fiber returned to routine use after years
of debate. We predict that over the next few years the term complex carbohydrates will also return on food labels as consensus over its definition continues
to be accepted.
As with any developing research, informed scientists must continue to
speak out about their findings and pursue answers to questions of analytical methods and physiological impact. Unlike marketing or advertising careers where professionals must keep pace with fads, most scientists and nutritionists have the
opportunity to stick with a subject until key issues are resolved and results show
that their work will either benefit mankind or must be set aside because it has
little or no value. The public understands how important complex carbohydrates
are. Health recommendations made by government agencies as well as those of
health and nutrition professionals have been pivotal in establishing this awareness.
The editors of this book seek to bring the issues that have been raised for
adding complex carbohydrates to the nutrition label closer to a solution. They
have also sought to bring health benefits and applications of complex carbohydrates to new food products development. This book includes international patent
activities on the use of complex carbohydrates (starch gums and dietary fiber)
in low fat and low calorie food products development as well as food content of
dietary fiber and carbohydrates. In all, it is hoped that a valuable view of the
current literature, practices, and prespectives is presented.
Susan Sungsoo Cho
Leon Prosky
Mark Dreher
Contents
Preface
Contributors
1. Introduction
Susan Sungsoo Cho, Leon Prosky, and Jonathan W. Devries
Part I: Health Benefits and Definition of Complex Carbohydrates
and Dietary Fiber
2. Dietary Guidelines for Complex Carbohydrates/Dietary Fiber
Joanne L. Slavin
3. Complex Carbohydrates and the Food Label: An FDA Perspective
F. Edward Scarbrough
4. Dietary Fiber Properties and Health Benefits of Non-Digestible
Oligosaccharides
M. B. Roberfroid
5. Suggested Alternatives to the Term Complex Carbohydrates
F. W. Scott, R. Mongeau, and P. Wood
11.
12.
Part III:
13.
14.
Part IV:
15.
16.
29.
30.
26
Roberfroid
Once they have reached the caeco-colon, most (but not necessarily all)
of the non-digestible oligosaccharides are hydrolyzed to small oligomers and
monomers that are further metabolized by one, a few, or most of the anaerobic
bacteria. Such a metabolic process, known as fermentation, not only serves the
bacteria by providing energy for proliferation, but also produces gases (H2, CO2,
CH4 ) which are metabolically useless to the host. Small organic acids such as
acetate, propionate, butyrate, and L-lactate, which are named short chain carboxylic acid (SCCA) are additional products. By a mechanism which is still a matter
of some debate (2, 3) the SCCA are largely (9095%) absorbed through the
intestinal wall, in the caecum and the ascending part of colon. Except for butyrate
(which is a major energy substrate for colonocytes) the other SCCA reach the
portal circulation and enter the liver, the main site of their metabolism. However,
part of acetate (2550%) escapes hepatic metabolism and, via the systemic circulation, reaches the peripheral tissues, mainly the muscle (4, 5). Even though they
do not provide the body with monosaccharides, the non-digestible oligo-/polysaccharides are, thus, indirect energy substrates and metabolic regulators. Thanks
to the symbiosis between the caeco-colon and a myriad of colonizing bacteria,
they are transformed to SCCA, which essentially via the same route as monosaccharides (portal blood), feed the liver and other important tissues with energy.
Acetate, propionate, butyrate, lactate, and so on are indeed important substrates
and/or intermediates in cell metabolism. Moreover, it has recently been demonstrated that, at least some of these molecules may also serve as modulators of
key metabolic pathways.
The most important class of dietary non-digestible saccharides is that composed of short (up to 10 monomers) and medium (up to 5060 units) chain-length
homopolymers (Table 1). Some NDOs, like inulin and its hydrolysis product
oligofructose, which belong to the group of fructans, are common natural food
ingredients (6) while others like galactooligosaccharides or Neosugar are synthetic products resulting from the enzymatic and/or chemical modification of
natural disaccharides like saccharose, lactose, and so on.
Among these NDOs the chicory fructooligosaccharides (inulin and its product of partial enzymatic hydrolysis, oligofructose) are authorized as food ingredients in most European countries. Others like trans-galactooligosaccharides (TOS
or GOS), isomaltooligosaccahrides (IMO), soybean oligosaccharides (SOS) and
xylooligosaccharides (XOS) are recognized as functional foods by Japanese authorities.
The aim of the present chapter is to review the nutritional and physiological
properties of chicory fructooligosaccharides that are, so far, the most studied
NDOs. Similar evidence exists, however, for the other NDOs. The objective is
to give scientific support to the classification of chicory fructooligosaccharides
as dietary fiber (7).
Non-Digestible Oligosaccharides
31
discussed concerns their caloric value. Indeed, being exclusive substrates for colonic fermentation, oligofructose and inulin have also much lower caloric value
than equivalent but digested carbohydrates. Moreover, they are (at least) partly
used as direct and indirect substrates for bacterial proliferation, a process that
further reduces their bioavailability to the host. Based on experimental evidence
as well as on basic biochemical knowledge a theoretical approach has been proposed to compare the metabolic efficiency (in term of ATP production) and thus
the effective caloric value of a fructosyl moiety in chicory fructooligosaccharide
and sucrose (as a typically digested substrate). As compared to sucrose chicory
fructooligosaccharide has a relative caloric value of 25 to 35%, or 1 to 1.5 kcal/
g or 4 to 6 kJ/g (16).
Mineral Bioavailability
As chicory fructooligosaccharides modify intestine and colon physiology, it
should be asked whether they might influence the nutritional balance of several
macro- or micronutrients.
Our recent studies (27) have shown that feeding rats with oligofructose or
inulin at the dose of 10% leads to an increased fecal excretion of N, resulting
most probably from an increase in fecal bacteria and leading to a decreased urinary excretion of urea and decreased uremia. Such results are in accordance with
those obtained with other dietary fibers.
The possible interference of dietary fibers with ion absorption relative to
their physiological effects on the small intestine and the colon has regularly been
questioned. In a recent experiment in rats fed a diet supplemented with 10% FOS
or inulin, we have demonstrated that the balance for Ca, Mg, and Fe is improved
(27). This beneficial effect, quite surprising as compared to the effect of dietary
fiber, may result from an increased intestinal absorption of the minerals. This
effect has also been observed by Levrat et al. (28) in rats fed a diet containing
10% inulin, and by Ohta et al. (29). All showed that in rats fed a diet enriched
in synthetic fructans the absorption of Ca, Mg and P was significantly
higher than in rats fed a diet supplemented with lactose. They also showed that
galactooligosaccharides had no effect. Neither Zn or Cu bioavailability nor lipid
soluble vitamins (A and E) homeostasis was modified by chicory fructooligosaccharides.
5
Suggested Alternatives to the Term
Complex Carbohydrates
F. W. SCOTT AND R. MONGEAU
Food Directorate, Nutrition Research Division, Banting Research Centre,
Ottawa, Ontario, Canada
P. WOOD
Centre for Food and Animal Research, Agriculture and Agri-Food Canada,
Ottawa, Ontario, Canada
INTRODUCTION
The Canadian Nutrition Recommendations published in 1990 by Health Canada
refer to the benefits of diets high in complex carbohydrates, defined essentially
as starches. For the purposes of food labeling claims in Canada the term complex
carbohydrates is permitted and has been taken to mean only starch which is the
term used on the nutrition label itself. In other jurisdictions, the term complex
carbohydrates has been taken to mean either starches or starches plus dietary
fiber. Thus, the term is a source of confusion, is difficult to define in a manner
that can be verified analytically and has little scientific meaning for many scientists and lay people alike (1, 2). There is a need to simplify and more clearly
define in chemical terms what is meant by the advice to consume foods high in
complex carbohydrates. It is time to re-think the use of this chemically undefined
7
The Role of Dietary Fiber in the
Prevention of Lipid Metabolism
Disorders
ELZBIETA BARTNIKOWSKA
Warsaw Agricultural University, Warsaw, Poland
INTRODUCTION
The earlier assumptions regarding the importance of dietary fiber relate to the
controversy on the differences of white and brown bread in terms of health and
better nutrition. Popular taste has tended to prefer white flour. Therefore white
bread with its higher cost and greater aesthetic appeal had social esteem, which
generally persisted until today.
The first researchers to pay attention to the health contribution of whole
cereal foods were S. Graham and J. H. Kellogg. History was not on their side
at that time, and the components of dietary fiber have largely been neglected as
they were considered to have no nutritive value. In the last three decades, however, attention has been focused on the fiber content in the diet. Interest in fiber
arose from epidemiological studies indicating the low prevalence of coronary
heart disease, diabetes, gallstones, obesity, appendicitis, diverticulosis, varicose
veins and hemorrhoids in underdeveloped countries, but common in Western
74
Cho et al.
Gravimetric Procedures
Crude Fiber Determination
The crude fiber (CF) method received official final action by AOAC in 1936 and
was officially approved for flour (AOAC method 920.86) and animal feed
(AOAC method 962.09) in 1920 and 1962, respectively (10). The technique involves digestion of the food sample with boiling, dilute acid and alkali with
corrections for residual ash content. Under these harsh conditions, all of the soluble fiber is lost with variable amounts of insoluble fiber leaving mainly lignin
and cellulose. Lignin and cellulose values are usually 1040% and 5090%,
respectively, of their original levels (3). Thus, the CF estimate underestimates
DF and does not give a constant value for any fiber component. Nevertheless,
CF determination is still used for nutritional labeling purposes in some countries
where Western diseases are not a major health concern.
78
Cho et al.
deaths would result in the United States if dietary goals for cancer prevention
such as increasing DF intake levels are not achieved (27). This could cost as
much as $25 billion a year. Based on international correlation statistics, an inverse
relationship has been found between fiber and fiber-containing foods and colon
cancer risk (28, 29). After examination of all fiber sources, the Life Science
Research Office (LSRO) (30) determined that the IDF-rich wheat bran most consistently reduced colon tumor incidence in animal models. Recurrence of precancerous polyp lesions in the rectum has also been shown to be lower with wheat
bran in humans (31). Both IDF and SDF may reduce breast cancer risk by binding
estrogen, a potent promoter, and thus preventing enterohepatic reabsorption and
lowering circulating levels (32). Although increased DF intake seems to be beneficial in terms of cancer, there are concerns about impaired mineral availability.
Examination of populations that consume much higher levels of DF (e.g. vegetarians vs. omnivores) showed that mineral levels in various biological samples were
comparable to those with lower DF intakes (30). In addition, bone mineral mass
was observed to be higher or at least the same. Thus, bioavailability of minerals
with higher DF intakes does not appear to be an issue as long as intakes are
adequate.
Other issues that are considered in making a recommendation are the intended audience, the current intake levels of DF, and how the DF will be consumed (i.e. in foods vs. supplements) (30). Recommendations are generally based
on a healthy, adult population and are not applicable to special populations like
young children and the elderly. In reviewing existing data on current DF intakes
of a population, the methods used for dietary assessment and chemical analysis
of the DF content of a food are determined since they both influence the estimated
intake level. Finally, recommendations are for DF in foods and not supplements.
The use of DF supplements may affect the balance of nutrients in an otherwise
healthy diet. Limited data exist on the effects of isolated DF which may differ
from the DF naturally present in a food (30).
81
(16) as suitable methodologies for the analysis of DF for nutrition labeling purposes. Professionals in the DF research field, however, strongly support the enzymatic-gravimetric technique as the most appropriate approach for nutrition labeling and quality control purposes (9). Thus, recommendations for DF intake in
both the United States and Canada are generally based on the AOAC methods.
Recommendations for DF intake in North America range from 25 to 35 g/d. The
U.S. FDA recommends that DF goals be based on a caloric basis (Table 2).
General dietary recommendations by organizations in the U.S. and Canada are
to increase intakes of grain products, fruits and vegetables which will increase
overall DF intake if followed (Table 4). Recent U.S. recommendations for intakes
of grain-based foods are at least 6 servings per day. In Canada, recommended
intake levels of breads, cereals and other grain products have increased from 3
5 servings/d to 512 servings/d (Table 4).
Even though pediatric guidelines for intakes in the U.S. have been set for
the consumption of other macronutrients such as protein, fat and carbohydrates,
no specific recommendations on DF levels have been made until recently. In
1993, the American Academy of Pediatrics (AAP) Committee on Nutrition made
the recommendation that DF intake in children over age 2 should be 0.5 g/kg
body weight (36) (Table 2). To avoid the potential hazards of excessive consumption, especially for overweight children, the AAP suggest a cap at 35 g/d. However, intakes up to 25 g/d should not be deleterious even with suboptimal mineral
intake (36). The American Health Foundation (AHF) recommends that a range
of DF intake, between Age 5 and Age 10 g/d may represent a safe
and tolerable level for most children over the age of 2 years (37) (Table 2).
Consistent with guidelines for adult DF intake, the AHF recommendation gradually increases the fiber intakes to the minimal adult DF level by age 20. Consumption of recommended levels of DF by children may help reduce the risk of developing chronic diseases such as cancer and cardiovascular disease later on in
life (37).
85
TABLE 4 Continued
Country
Canada
Canada
Name/Source of recommendation
Canadian Cancer Society, Fact on
Cancer and DietYour Food
Choices May Help You Reduce
Your Cancer Risk (1986)
Health and Welfare Canada, Canadas Food Guide Handbook
(1985)
89
the U.S. to demonstrate relative proportions in which different food groups should
be consumed (Table 6). Recommendations are to emphasize grain-based foods
like rice, breads, and cereals.
Recommendations for DF intake are lower in Japan (2025 g/d) than in
Australia (30 g/d) (Table 2). Australians have dietary habits similar to that of
Americans. However, officials in Japan believe that the American RDA for DF
is probably too high for the Japanese considering the lower energy consumption,
lower fat intake, and insufficient calcium intakes which may be exacerbated by
high DF levels (40). Another point for consideration was the recent fad in Japan
to consume drinks containing 5 g/bottle or more of soluble oligosaccharides and
the unknown health impact if DF recommendations were set at a higher level.
In determining appropriate DF intakes for Japanese, Nishimune and colleagues
estimated the TDF intakes in 1935, 1955, and 1990 using a database with values
derived from the AOAC enzymatic-gravimetric method and, using the same
method, measured the TDF of model duplicate meals and composites for the
average Japanese in 1985 (40). Using the average Japanese energy intakes of
2,088 kcal in 1985, suggested daily DF intake levels are 1012 g/1,000 kcal
(about 2025 g/d). The lower limit is based on the DF required for a daily fecal
output of 140150 g since this is associated with bowel disease risk (41). Factors
considered in establishing the upper limit are mineral sufficiency, fat consumption, ratio of IDF to SDF intake, the decreasing trend in Japanese TDF intake,
and the increase in mortality rates of diabetes mellitus, ischemic heart disease,
and diverticulosis.
92
Cho et al.
isolated DF products, they also state that it is preferable to consume foods naturally high in fiber than to add bran to low fiber foods. Increasing fiber intake
by consuming unrefined cereal products like brown and whole wheat bread and
high-fiber breakfast cereals is advocated. The recommended DF intake is 30-40
g TDF/d (Table 2).
94
Cho et al.
as a conversion factor (4850). Since the CF level can vary from 1050% of
the TDF value depending on the nature of the food, dietary intake data employing
the CF method should be interpreted with caution. Tables of DF levels in foods
have been improved by employing AOAC TDF 985.29/991.43, NSP, or unavailable CHO methodology, but limited intake data are available using these new
databases.
102
Cho et al.
the NSP contribution from cereals greatly increases the overall NSP intake (62).
Unfortunately, negative trends in DF consumption have been observed in some
countries. Dietary fiber intakes in Japan have been declining consistently since
the 1950s (40, 63). Similar drops are being observed in certain rural and urban
areas of Latin America (50, 64). So although some positive dietary patterns have
been observed in certain regions (52, 65), especially in Italy where significant
positive trends in DF intake have been observed (66), dietary intake levels of
DF are still falling short of the recommended levels.
Dietary fiber intake levels can also vary according to sex, ethnicity, and
lifestyle. Women usually have lower intakes than men. In some cases, this is due
to lower energy intakes. Some investigators have found that although womens
overall intake of DF was lower, they chose more fiber-dense diets (57) consisting
of foods such as brown breads, cereals, raw fruit, and salads (54). However, men
and younger people tended to consume more DF from potatoes cooked in fat
and pulses in one study (54). In the U.S., Block and Lanza found trends in DF
intakes also differ between the sexes. Dietary fiber intakes in women increased
with age whereas the opposite occurred in men (67). In the same study, ethnic
differences in DF intakes existed with Blacks tending to consume less than
Whites in both sexes across all age groups (67). However, Ballew and Sugerman
found that DF intakes in Mexican women living in Chicago were different than
those living in Texas, suggesting that within an ethnic group there are also temporal and regional variations in dietary intake patterns (68). Greater DF intakes in
rural vs. urban areas is not uncommon (69). Lifestyle plays a large role in the
level of DF consumed. In general, those with healthier lifestyles or concerned
about getting adequate nutrient intakes had higher intakes of DF (53, 7073).
As in the adult population, DF intakes in children are lower than recommended levels, but their dietary patterns are also undesirable. In the U.S., DF
intakes of children are not increasing with age according to the AHF Age
5 guideline (74). Mueller and colleagues found that as children age, their consumption of fruits, vegetables, and cereals decline (74). Dietary fiber intake levels
are lowest in Japanese under the age of 19 (40). In the Child and Adolescent
Trial for Cardiovascular Health (CATCH) study (Table 10), dietary intervention
to reduce total fat in school children did indicate a shift in energy from fat to
CHO while keeping energy level from protein stable. However, the relative proportions of energy from simple sugars and complex CHO remained the same
(75). Norwegian adolescents consume too much fat and sugar and not enough
DF (70). In Sweden, young people may have adopted advice about consuming
less fat, but instead of increasing their fiber intake, consumption of sweets and
soft drinks is considerable (76). Thus, it is important to educate the youth about
the importance of consuming adequate amounts of dietary fiber as part of a
healthy diet.
Adequate intakes of DF are necessary to prevent the onset of constipation
117
(available), resistant starch, cellulose, hemicelluloses, pectic substances, hydrocolloids (gums, mucilages), modified starches, and resistant oligosaccharides.
There is much yet to be learned about this area and the components involved,
e.g. isolation, structures, properties, physiological responses, definitive analytical
procedures. It is made difficult by the fact that complex carbohydrates cannot be
clearly defined on the basis of structure, analytical methodology, or physiological
response. The components of complex carbohydrates/dietary fiber have traditionally been defined on the basis of the solvent extraction procedures used for their
isolation. This does not result in clearly definitive separations of the components,
nor in definition when a structural basis is used.
STARCH
Starch is the most important reserve polysaccharide of many higher plants where
it occurs as discrete granules in the leaf, stem (pith), root (tuber), seed, fruit and
pollen. The size, shape and gelatinization temperature of the granules depend on
the botanical source of the starch. Common food starches are derived from seed
(wheat, maize, rice, barley) and root (potato, cassava/tapioca) sources.
Starch consists of a mixture of two polymers, amylose and amylopectin,
that are composed entirely of glucose units, i.e. homopolysaccharides. The glucopyranosyl units in amylose are linked through alpha-D-(1 4)-glucosidic linkages. Amylose has traditionally been considered to be a linear polymer with an
average chain length and degree of polymerization of approximately 1000 or less.
However, it is now known that it contains a limited amount of branching involving alpha-D-(1 6)-glucosidic linkages at the branch points. Amyloses, as isolated, are often a mixture of linear and branched molecules (Figure 1). The behavior of amylose is dominated by two major properties:
(a) Its ability to form inclusion complexes with iodine, aliphatic primary
alcohols, lipids and surfactants. Amylose produces a deep blue inclusion complex
with iodine, used in the detection of starch, in which the amylose chain forms a
helix around the iodine molecule with six glucosyl units per turn of the helix.
(b) Its ability to form strong intermolecular interactions, hydrogen bonding,
leading to precipitation or gel formation when a solution of amylose is cooled.
Amylopectin is a high molecular weight, highly branched polymer containing about 56% alpha-D-(1 6)-glucosidic linkages. It has an average chain
length of 20 to 25, an average degree of polymerization in the thousands, and
molecular weights in the millions. The structure of amylopectin contains clusters
in which about 2025 chains containing 1216 glucose units are involved (8)
(Figure 1). These clusters, in which many of the chains exist as double helices,
are responsible for the crystalline regions of starch granules.
While the manner in which amylose and amylopectin are organized to form
the granule is not clearly understood, the granule is partially crystalline exhibiting
120
Lineback
celluloses were those polysaccharides solubilized by aqueous alkali after watersoluble and pectic substances were removed (10). The situation is actually much
more complex. Cell wall polysaccharides exist as heterogeneous mixtures of
polysaccharides that are associated by hydrogen bonding and even covalently
linked to varying extents. These include homopolysaccharides, those that contain
only one type of carbohydrate monomer such as cellulose and mixed-linked betaglucans, and heteropolysaccharides, those that contain more than one type of
carbohydrate monomer such as arabinoxylans, galactomannans and xyloglucans.
Structural determination and characterization of these polysaccharides are
made difficult by several factors. For each carbohydrate monomer involved, the
following must be determined (11):
1.
2.
3.
4.
5.
6.
Hemicelluloses may be present in soluble and insoluble forms and are comprised of a number of branched and linear pentose- and hexose-containing polysaccharides. In cereal grains, soluble hemicelluloses are termed pentosans.
Hemicelluloses are of much lower molecular weight than cellulose. Component
monosaccharide units may include xylose, arabinose, galactose, mannose, glucose, glucuronic acid and galacturonic acid.
Hemicelluloses are normally classified by the predominant carbohydrate
monomer present. Illustrations are given in Table 1. The most abundant and widespread noncellulosic cell wall components are the xylans, polymers with a main
chain of beta-D-(1 4)-linked D-xylose. The backbone structure may be substituted with single (4-O-methyl)-alpha-D-glucopyranosyluronic acid units, single
alpha-L-arabinofuranose units or longer side chains in which the L-arabinofuranose residues have additional substituents. A general structure for xylans is shown
below: some side chains (R) include D-galactose, L-galactose, and D-xylose
units.
123
The structures of furcellaran and alginates also contain sulfate esters. Three carrageenan fractions exist: one (kappa) does not gel, one (lambda) forms brittle gels
in the presence of potassium ions, and one (iota) yields a more elastic gel in the
presence of calcium ions. Alginates are linear copolymers of D-mannuronic acid
(M-blocks) and L-guluronic acid (G-blocks) with regions (blocks) containing
only one of the acids. Solubility properties are controlled by the ratio of mannuronic acid blocks to guluronic acid blocks. Alginate gels can be formed in the
cold in the presence of calcium which induces intermolecular associations in the
G-block regions and have sufficient stability to withstand heat processing.
Gums arabic, karaya and tragacanth are complex heteropolysaccharides derived from plant exudates. Guar and locust bean (carob) gums, obtained from
seeds, are galactomannans where the mannan is made soluble by single galactose
unit side chains. Guar gum (mannose to galactose ratio of 2: 1) is more substituted
than locust bean gum (4 :1 ratio), and is cold water soluble. Locust bean gum
requires hot water. An example of a mucilage not commercially available but
used to thicken foods during cooking, particularly in the southeastern part of the
United States, is that from okra. This is a rhamnogalacturonan in which L-rhamnose is linked to D-galacturonic acid units in the interior chain of the polymer.
Xanthan gum, produced by microbial fermentation, has a cellulose main
chain that is made soluble by the presence of short side chains on every second
glucose unit. These side chains contain glucose, mannose, and glucuronic acid
units in a 3 :3 :2 ratio. Xanthan has a molecular weight in the millions and forms
a synergistic gel with locust bean gum but not with guar gum.
While these are used in small amounts in food products, their complex
carbohydrate nature and structure cause them to function as a component of dietary fiber. They are not digestible by the enzymes of the human gastrointestinal
system.
MODIFIED STARCHES
Many starches do not have the functional properties needed to impart or maintain
desired qualities in food products. As a result, starches have been modified to
obtain the functional properties required. The types of modified food starches,
also known as starch derivatives, are listed in Table 2 (16).
The starches most commonly modified for commercial use are those from
normal maize, tapioca, potato, and waxy maize. The methods used to produce
modified starches include dextrinization, acid-hydrolysis, oxidation, cross-linking, monosubstitution by esterification and etherification, and combinations of
these processes. The manufacture of food starches (chemicals used, amount incorporated into the starch, amount of residues permitted) is regulated by the Food
and Drug Administration (FDA) and listed in the Code of Federal Regulations
(21CFR 172.892). Modified food starches must be approved by the FDA as food
Complex Carbohydrates
133
No. of
persons
No. of
countries
58
31
21
4
18
2
5
4
134
No.
N 107
Percent
77
74
62
62
35
34
31
72%
69%
58%
58%
33%
32%
29%
Available starches
Non-starch polysaccharides (NSP)
Total starches
Unavailable starches (Resistant starches)
Oligosaccharide, available
Oligosaccharide, unavailable (Resistant oligosaccharides)
Lignin
Complex Carbohydrates
135
Definition provided
Polysaccharides
Glucose polymers which are slowly digested or not
digested in the small intestine
Carbohydrates minus simple sugars
Dont like the term
Starch and dietary fiber
Available starches
Starch dietary fiber oligosaccharides
Glucans with a complex structure
Dietary fiber unavailable oligosaccharides
NSP lignin
Starch and cell wall carbohydrates
Unavailable carbohydrates
Glucans not soluble in 80% ETOH
Carbohydrates of plant origin (DP4)
No. of
respondents
providing the
definition
Percent
19
10
26%
14%
10
9
7
4
4
2
1
1
1
1
1
1
13%
9%
5%
5%
5%
3%
1%
1%
1%
1%
1%
1%
Complex Carbohydrates
139
Flow Rate:
1.0 mL/min.
Injection Temperature:
room temp.
Elution Mode:
Isocratic
Detector:
Refractive index
Detector Sensitivity:
0.020 x 103
Response Time:
2.0 min.
Run Time:
25 min.
Injection Volume:
15 L
Note: Parameters may vary in order to optimize the chromatography.
sugar extracted, %
100
sugars in samples, %
Analytical Results
The HPLC method described in this paper is a quantitative method for available
starch in foods. A variety of samples, such as grains, ready-to-eat breakfast cereals, breads, crackers, fruits, vegetables, and mixed diets can be analyzed by using
this method. Complex carbohydrates are determined as the sum of available
starch and dietary fiber; available starch by high performance liquid chromatography determination of glucose and maltose present in the enzymatic hydrolysates
of desugared sample; and dietary fiber by enzymatic gravimetric method, official
final action method 991.43.
It is important to remove simple sugars prior to enzymatic hydrolysis of
starches, so that simple sugars do not carry over in the sugar analysis for available
starch determination. For the efficient desugaring, we used 49% ethanol in water
initially for easier dissolution of sugars. Additional 95% ethanol was added to
make the final concentration of 78% ethanol to precipitate dietary fiber extraction.
This procedure would ensure that simple sugars are efficiently removed without
eliminating water soluble dietary fiber components. Table 5 indicates that the
extraction efficiency is generally excellent, ranging from 90% to 99%. One cereal
sample which showed 90% desugaring efficiency had a low sugar content (6.2%)
in the original sample to begin with. Ethanolic extract of that sample contained
5.6% sugars. We consider both 6.2% and 5.6% to be in the normal analytical
variability range for sugar determinations. Thus, overall efficiency of the desugaring step is considered excellent.
The elution orders of standard sugars in this HPLC method, using an aminobonded silica column packing and acetonitrile-water mobile phase, follow the
expected saccharide polarity: fructose, glucose, sucrose, and maltose. The enzy-
140
Source
Standard Mix
Green Bean
Wheat Bran Cereal
Prune
Bread
Cellulose
Wheat Starch
Cracker
Sugar extracted,
%
Sugars in
sample,
%
% Extraction
efficiency
39.8
15.6
15.9
53.5
5.6
0
0
1.0
40.0
16.2
17.0
56.2
6.2
0
0
1.0
99.0
96.3
92.9
95.2
90.3
0
0
0
matic digestate of the desugared sample showed mostly glucose peaks and negligible peaks from maltose and sucrose. The sample chromatograms demonstrated
that the starch digestion steps using three enzymes, heat stable alpha-amylase,
protease, and amyloglucosidase, were near completion, producing mostly glucose. The glucose values in the enzyme hydrolysate were multiplied by the factor
0.9 to convert into starch values.
The Final Food Labeling Regulations in the United States (6) have recognized that HPLC methods are the most appropriate for nutrition labeling purposes among all the sugar determination techniques. The HPLC approach
offers the most accurate and specific determinations of mono- and di-saccharides. Thus, the HPLC technique can give the most accurate figures for available
starch, which is derived from glucose and maltose values of the starch hydrolysates. However, the method requires a high capital investment and trained personnel for troubleshooting. The presence of salt, especially in the hydrolysate of
processed foods, interferes with the analysis because chloride ions elute quickly
after glucose. It is important to wash the column for 2 hours at 1.5 mL/min with
a solution of 0.1% tetraethylenepentamine (TEPA) (pH approximately 7 with
acetic acid) in 78% acetonitrile in water and finally with normal mobile phase
(20).
Table 6 shows the percent recovery of several standards. Percent recoveries
of available starch and DF for these known standards are excellent. The starch
digestion in our method was done by heat stable alpha-amylase and amyloglucosidase, which are more specific to glucose-glucose bonds. Thus, most of the sucrose
in the sample (99%) was recovered as sucrose in the enzyme hydrolysate.
In Table 7 are the reported available starch data generated by HPLC and
146
Li
METHOD
Triplicate samples (0.5 g) of freeze-dried foods are weighed into teflon tubes and
extracted with 15 mL of n-hexane if the samples contain 5% fat; residues are
ex-tracted with 1015 mL of 80% methanol (Figure 1). The alcohol extract is
used for the separation and quantitation of individual sugars using GLC (Figure
2). Residues after the alcohol extraction are suspended in 25 mL deionized water
and autoclaved at 121C for 1 h, cooled and mixed with a solution of amyloglucosidase (No. 208-469, Boehringer-Mannheim Corp.) in acetate buffer; the mixture is incubated at 55C for 2 h. Aliquots (0.1 mL) of the hydrolysate are removed, dried, and derivatized for GLC analysis for glucose. Starch content is
calculated as glucose 0.9 (Figure 3). To the remaining hydrolysate and suspensions is added 4 volume of 95% ethanol. After 1 h, the mixtures are filtered
through glass crucibles matted with Celite (CAFA); residues are dried at 105C
and weighed. One of the triplicate residues is analyzed for crude protein as nitrogen 6.25, the second is heated in a muffle furnace at 525C for 5 h for ash
determination (Figure 4). The third is hydrolyzed in H 2 SO 4; aliquot of the acid
hydrolysate is derivatized to alditol acetates and analyzed for neutral sugars by
GLC, and for uronic acid colorimetrically (Figure 5) (2). Total dietary fiber (TDF)
is the weight of residue after enzyme hydrolysis corrected for the weights of
crude protein and ash. Dietary fiber polysaccharides (DFP) is the sum of neutral
sugars and uronic acid in the residue.
Carbohydrate Fractions
149
residue obtained using either one of the AOAC methods, the determination of
nitrogen for crude protein correction is especially critical with this method. However, the method is less labor intensive and more cost effective. The quantitation
of glucose in starch hydrolysate can be done using GLC, HPLC, or other enzymatic methods. The above method was used to analyze a number of frequently
consumed foods that had been stored at 20C after they had been analyzed for
sugars by HPLC, soluble and insoluble dietary fiber by AOAC method 991.43.
The earlier analyses were done in a commercial laboratory (4) with a contract
from the Nutrient Data Laboratory, ARS, USDA (formerly the Human Nutrition
Information Service, USDA). Detailed procedures of our method and comparisons between the two laboratories of the data on sugars and dietary fiber contents
of the same 34 foods are given elsewhere (3). Sample data from our laboratory
on total sugar, starches, and total dietary fiber are presented here for 4 groups
of foods. Table 1 gives the values for baked products; Table 2 for cereal grains
and pasta; Table 3 for legumes, and Table 4 for vegetables. Table 5 gives the
values for individual sugars, neutral sugars [(arabinose fucose galactose
glucose mannose rhamnose xylose) 0.89], uronic acid, dietary fiber
polysaccharides (neutral sugars uronic acid) and total dietary fiber of 12 selected foods.
Based on the dry weight of selected foods, baked products contain between
50 and 70 g/100 g of starch, 6 and 18 g/100 g of TDF; cereal grains and pastas
contain between 55 and 90 g/100 g of starch, 2 and 10 g/100 g of TDF; canned
legumes contain between 20 and 40 g/100 g of starch, 20 and 30 g/100 g of
Carbohydrate Fractions
151
Total sugar
0.55
1.14
0.54
0.82
Starch
89.33
74.90
67.20
72.55
55.50
0.03
0.05
0.03
0.01
0.51
7.90
2.12
1.34
1.55
1.79
5.08
4.86
8.36
10.73
0.06
0.03
0.05
0.01
0.12
Sugar
14.18
6.27
1.59
1.70
1.66
Total starch
0.12
0.15
0.02
0.05
0.09
18.58
42.00
27.81
33.11
34.18
1.32
0.17
1.07
1.09
2.45
0.63
0.75
0.33
0.71
0.62
Sugar
28.48
1.20
21.89
18.62
0.24
0.12
0.23
0.38
25.34 0.33
Total starch
4.74
77.12
23.32
46.90
0.12
0.63
0.47
1.35
15.02 0.04
0.24
0.14
0.44
0.67
18.01 0.74
160
Champ et al.
Intake (g/d)
Starch
NSP
Ref.
380
380
190
208
95.5
96.7
83.7
96.9
94.8
98.6
35
41
28
25
50
62
(6)
(6)
(7)
(7)
(8)
(8)
QUANTIFICATION OF RS IN HUMANS
One indirect and two direct methods have been used to estimate RS in humans.
They are presented in Table 7.
H2 Breath Test
Hydrogen is one of the end-products of carbohydrates fermentation. It is exclusively formed in the colon by the bacterial fermentation, partly absorbed and
In Vivo Techniques
161
Shortcomings
Efficiency of Nebacitin
Endogenous -amylase in the large intestine
Shortcomings
Requires specific equipments
High cost
Correlation with data/humans?
H2 excretion in rats
Measurement of total H2 excretion (breath and flatus) in a respiration chamber
Advantages
Non-invasive
Shortcomings
Requires specific equipments
Not suited as a quantitative model
Shortcomings
Requires skilled persons to perform surgery
Microflora not completely eliminated (Cc)
Digestibility of starch overestimated (Cc)
cleared in a single passage of the lungs, then excreted in the expired lungs. The
results of gas perfusion techniques have suggested that a rather constant fraction
of the total H2 production is excreted by the lungs, and that rates of breath H2 and
H2 production correlate well (15). Using a non-absorbable but quickly fermented
oligosaccharide (the lactulose) to calibrate the subject, it can be expected to
quantify a malabsorbed carbohydrate. Whereas this procedure remains quantitative for oligosaccharides it is only qualitative for insoluble or slowly fermented
substrates. Table 8 illustrates the finding that in many cases, the method is inappropriate to quantify RS. The comparison of the RS values obtained by the H2
breath test and by intubation of healthy subjects (direct measurement) when the
168
Champ et al.
13. Bach Knudsen, K. E., Agergaard, N., Olesen, H. P. (1991) J. Anim. Physiol. Anim.
Nutr. 66, 190203.
14. Hildenbrandt, L. A., Marlett J. A. (1991) J. Nutr. 121, 679686.
15. Levitt, M. D. (1969) N. Engl. J. Med. 284, 13941398.
16. Bornet, F., Cloarec, D., Gouilloud, S., Champ, M., Colonna, P., Barry, J. L., Galmiche, J. P. (1990) Gastroenterol. Clin. Biol. 14, A90.
17. Scheppach, W, Bach, M., Bartram, P., Christl, S., Bergthaller, W., Kasper, H. (1991)
Dig. Dis. Sci. 36, 16011605.
18. Wolever, T. M. S., Cohen, Z., Thompson, L. U., Thorne, M. J., Jenkins M. J. A.,
Prokipchuk, E. J., Jenkins, D. J. A. (1986) Am. J. Gastroenterol. 81, 115122.
19. Levitt, M. D., Hirsch, P., Fetzer, C. A., Sheahan, M., Levine, A. S. (1987) Gastroenterol. 92, 383389.
20. Lund, E. K., Johnson, I. T. (1991) J. Nutr. 121, 311317.
21. Christl, S. U., Murgatroyd, P. R., Gibson, G. R., Cummings, J. H. (1992) Gastroenterol. 102, 12691277.
22. Stephen, A. M., Haddad, A. C., Phillips, S. F. (1983) Gastroenterol. 85, 589595.
23. Sandberg, A. S., Andersson, H., Hallgren, B., Hasselblad, K., Isaksson, I. (1981) J.
Nutr. 45, 283294.
24. Sandberg, A. S., Andersson, H., Kivists, B., Sandstrsm, B. (1986) Br. J. Nutr. 55,
245254.
25. Englyst, H. N., Cummings, J. H. (1985) Am. J. Clin. Nutr. 42, 778787.
26. Englyst, H. N., Cummings, J. H. (1986) Am. J. Clin. Nutr. 44, 4250.
27. Molis, C., Champ, M., Flouri, B., Pellier, P., Bornet, F., Colonna, P., Kozlowski,
F., Rambaud, J. C., Galmiche, J. P. (1992) Eur. J. Clin. Nutr. 46, S131S132.
28. Faisant, N., Bulon, A., Colonna, P., Molis, C., Lartigue, S., Galmiche, J. P., Champ,
M. (1995) Br. J. Nutr. 73, 111123.
29. Flouri, B., Leblond, A., Florent, F., Rautureau, M., Bissalli, A., Rambaud, J. C.
(1988) Gastroenterol. 95, 356363.
30. Noah, L., Guillon, F., Bouchet, B., Bulon, A., Gallant, D. J., Colonna, P., Molis,
C., Faisant, N., Galmiche, J. P., Champ, M. (1995) in Proc. Eur. Assoc. for Grain
Legume Research Conf. July 913, pp. 276277.
31. Bornet, F. R. J., Cloarec, D., Barry, J. L., Colonna, P., Gouilloud, S., Delort-Laval,
J., Galmiche, J. P. (1990) Am. J. Clin. Nutr. 51, 421427.
32. Rumessen, J. J. (1992) Eur. J. Clin. Nutr., 46(S2), S77S90.
33. Andersson, H. (1992) Eur. J. Clin. Nutr., 46(S2), S69S76.
34. Langkilde, A. M., Andersson, H., Faisant, N., Champ, M. (1995) in Proceedings of
the concluding plenary meeting of EURESTA, N. G. Asp, J. M. M. van Amelsvoort & J. G. A. J. Hautvast (Eds), European Flair-Concerted Action on Resistant
StarchNo. 11 (COST 911), pp. 2830.
35. Mougham, P. J., Cranwell, Darragh, A. J., Rowan, A. M. (1994) in Proc. VIth International Symposium on Digestive Physiology in Pigs. Souffrant W.-B. & Hagemeister H. (Eds), Bad Doberan, D, 46 October 1994. pp. 389396.
Analytical Methods
173
into account the potentially digestible starch (oligosaccharides and part of the
high molecular weight fraction) (7,8).
General Principle
In order to quantify RS, the first step is to remove the digestible starch from the
sample. This is performed using a pancreatic (-amylase. In some of the methods,
an amyloglucosidase is added in order to avoid a possible inhibition of the amylase by the products of the digestion (mainly maltose, maltotriose). The amylolysis is also sometimes preceded by a proteolysis which is supposed to reflect
the action of the pepsin inside the stomach and of the trypsin which is secreted
in the pancreatic juice together with the -amylase.
After the hydrolysis of the digestible starch, RS is quantified directly in
the residue (isolated most of the time by 80% ethanolic precipitation) (6,9) or
by difference between total starch and digestible starch which are quantified separately (2).
Analytical Methods
175
Procedure (Figure 3)
Main Characteristics
Fresh foods can be analyzed as eaten. They are passed through a mincer.
The hydrolysis is performed in presence of glass ball and guar gum (to
prevent mechanical alteration of very sensitive starch such as raw potato
starch).
The samples are hydrolyzed with pancreatin, amyloglucosidase and invertase. The pancreatin contains several enzymic activities including
-amylase and proteolytic enzymes which allows the digestion of the
digestible material (starch and proteins). In contrast, the lipolytic activity of the pancreatin might be ineffective due to the absence of bile in
the medium, into glucose and fructose. The invertase is added in order
to hydrolyze all the sucrose present in the sample. The sucrose must
then be quantified separately. This step has been introduced in the
method because of the presence of an invertase contaminating activity
in the amyloglucosidase. This activity could lead to an overestimation
of the starch content of the sample.
The digestible starches (starches hydrolyzed after 20 and 120 min hydrolysis) are quantified on an ethanolic extract (64.4% ethanol). The
extract is hydrolyzed into glucose by amyloglucosidase. Then the glucose is quantified using GOD-PAP kit.
RS content is determined by difference between total starch and digestible starch contents.
Analytical Methods
177
The Method of Muir & ODea, 1992 & 1993 (14, 15)
Principle
The chewed food sample is incubated with pepsin, then hydrolyzed by -amylase
and amyloglucosidase. The insoluble residue represents RS.
Procedure (Figure 4)
Main Characteristics
The food is first chewed by a volunteer in standardized conditions. The
sample weighed for the analysis contains about 100 mg carbohydrate.
Pepsin, -amylase and amyloglucosidase are used for the hydrolysis of
the digestible fraction of the sample.
RS is recovered by centrifugation followed by aqueous washing of the
pellet.
Starch in the residue is hydrolyzed into glucose by Termamyl then pancreatin and amyloglucosidase. Dimethylsulfoxide is used prior to the
second hydrolysis step to allow a complete dispersion of the RS. Glucose is then quantified using a GOD-PAP kit.
Advantages and Drawbacks
This method has been validated with in vivo studies on ileostomates (15,16).
From the 7 comparisons available, it seems satisfactory. No comparison with
other in vitro have been published. Being used, until now, only by the authors,
difficulties and reproducibility cannot be evaluated.
The Method of Champ et al., 1992 (6)
Principle
In vitro RS was defined as the starch not hydrolyzed by incubation with amylase. Hydrolysis products are extracted in 80% ethanol and discarded. RS is
then solubilized with 2N KOH and hydrolyzed with amyloglucosidase. In the
new procedure, amyloglucosidase is added to the pancreatic -amylase to avoid
inhibition of the amylase by the products of the digestion.
Procedure (Figure 5)
Main Characteristics
In the first two methods, the sample has to be dried (freeze-dried when
possible) and ground into fine particles. In the new procedure, fresh
Analytical Methods
181
In vivo
In vitro
(10)(20 meth. C)
Ref.
29.2
4.5
18.1
4.1
32.5
10
8
11
23.429.0
7.09.2
13.917.5
4.2
32.2
8.7
11.9
8.3
(20)
(20)
(20)
(21)
(22)
(22)
(22)
(22)
Fed (g)
Recovered (g)
Mean
recovery (%)
0
8.5
8.5
11.7
15
0.3
9.0
8.6
13.7
13.7
106
101
117
91
196
Ouarne et al.
was obtained on the integrator for concentration values ranging from 0.15 to 70
mg/L. The lower detection limit was based on a peak height at least three times
that of the background noise.
FIGURE 1
197
small cake (J. Pasquier patissier madeleine superstar) after FOS addition
showed glucose (G), fructose (F), sucrose (GF), GF2, GF3 and Rs1 and Rs2.
To investigate the nature of the oligosaccharides present in cakes containing
FOS, the supernatants obtained during sugar extraction were incubated in the
presence of invertase.
After the invertase action (Figure 3) the disappearance of oligosaccharides
can be observed, correlated to an increase in glucose and fructose concentrations.
On the other hand, Rs1 and Rs2 were not hydrolyzed, and appear on the chromatogram. If the specific areas of GF2 and GF3 were related to Rs1 and Rs2,
respectively, the theoretical concentrations of these sugars in the supernatants
analyzed could then be determined. The theoretical amounts (in g) of Rs1 and
Rs2 in cakes can be calculated from the chromatogram corresponding to Figure
3. The recovery is then calculated as follows:
[(GF2 RS1) RS1] [(GF3 RS2) RS2] GF4
100
(GF2i GF3i GF4i)
where:
199
Plain yogurt
Bio yogurt
Fruit yogurt
97.4
99.1
97.6
98.0
100.3
103.9
100.9
99.8
98.9
101.2
99.7
1.9
100.4
102.6
97.8
97.5
102.1
98.1
99.8
94.4
98.4
101.4
99.2
2.4
101.1
99.9
100.4
99.7
100.0
97.7
99.0
100.0
98.3
96.4
99.2
1.3
201
REFERENCES
1.
2.
3.
4.
5.
204
Coussement
method was elaborated. This was based on the use of a specific inulinase enzyme,
resulting in the total elimination of all fructans from the AOAC TDF determination.
An overview of analytical methods that can be used for the determination
of fructans is presented. The general method which can be used in all circumstances can be called the inulinase method for fructan determination.
Inulin (%)
Oligofructose (%)
16
210
316
911
410
29
14
0.30.7
1620
813
319
1520
14
26
25
36
25
1
23
0.30.7
1620
510
209
Direct HPLC
Direct HPLC (2) can be used reliably for quantitative determination of oligofructose, but not for inulin except for pure samples. The procedures are relatively
simple and straightforward. The DP information is not very reliable: peaks do
not represent one single DP fraction (example: the F4 fraction mixes with the
GF2 fraction) and from DP 4 the accuracy and separation are low. There can be
significant interferences with other oligosaccharides (maltodextrins, galactooligosaccharides, others) and other organic components.
Direct GC
Although these methods are more complicated, they provide the most accurate
information on the oligosaccharide fraction (2). They cannot be used for inulin.
These methods provide well separated chromatographic peaks for the GFn and
Fn fractions, thus allowing their accurate measurement up to about DP 10. There
are almost no interferences with other oligosaccharides, allowing the differentiation of different types of oligosaccharides in the same mixture. The sample preparation can be standardized to such an extent that very accurate and reliable measurement is possible.
HPAEC/PAD Methods
These methods, although providing interesting qualitative results, are difficult to
use for quantitative determinations. However, also here significant progress has
been achieved recently (non-published results).
210
Coussement
are known, which is not often the case in foodstuffs. Therefore, these methods
are mainly used for production control reasons.
Inulinase Method
This method (Fig. 3) allows the accurate determination of the total fructan content
of any nature in foodstuffs (12). The principle of the method is the specific hydrolysis of fructans using an inulinase enzyme. The calculation is done based on a
comparison of the glucose, fructose and sucrose determinations before and after
hydrolysis. The analysis of these mono- and disaccharides can be done by any
reliable method (not with enzymatic methods). To avoid interference with, e.g.,
lactoses, maltitol and isomalt a GC method is preferred.
The method has been used in many laboratories for quite some time, and
has been perfected. In our experience, it has proven to be reliable and accurate
in practice. A more thorough inter-laboratory comparison is being prepared now.
The inulinase method does not allow one to distinguish between inulin and
oligofructose, and does not give information on DP fractions. The method is
accurate in all food samples and requires no significant sample pretreatment.
There are almost no interferences with other substances.
The method can be combined with the modified AOAC method as described in Chapter 5 (Figures 4 and 5). It can also be combined with any of the
other methods described above to allow the differentiation between inulin and
oligofructose.
FIGURE 3
215
(5) wherein the soluble fibers precipitate at 60C in ethanol. Because inulin has
a relatively low average molecular weight (MW 6,000) as compared to the traditional soluble fiber (MW 10,000 to 50,000) it does not precipitate completely
under these conditions.
219
% -FDF
where:
R1 concentration of the total fructose (g/l)
R2 concentration of the free fructose (g/l)
R3 concentration of the fructose (g/l) from sucrose
P mean mass (g/l) of the duplicate test samples,where P 5 times the
mean dry mass of the test sample taken
Empirical conversion factor for fructose to fructan, assuming a ratio of 80 to
85% fructose and 15% glucose in the fructan polymer.
Modified A.O.A.C. Dietary Fiber Method 991.43 Without -Fructan
Dietary Fiber
As shown in Figure 2, we apply A.O.A.C. TDF method 991.43 (Lee et al., Ref.
5), but with some modification. This minor modification includes only a stage
of inulinase digestion performed, after the amyloglucosidase digestion. It aims
to eliminate the fructans, under the following conditions:
inulinase: Novozym SP 230 1800 inu/g, 100 ml
temperature: 60C; pH: 4.5
The pH adjustment step for the inulinase is the same as for amyloglucosidase.
Results
Inulin AssaySamples Without Sugar
Inulin samples were digested with Novozym SP 230 to check the performance
of the hydrolysis according to the HPLC procedure for samples without sugar.
As shown in Table 1, the enzymatic digestion recovery is higher than 95%.
The moisture contents of all the samples, as determined separately, were
ca 5%. The correction factor applied to compute the amount of inulin in the
sample varies between 1.0 and 1.2 for the different fructans. However, in practice,
a correction factor 1.1 for routine analyses should procure fairly accurate results.
222
Dysseler et al.
cooled and made up to 100 ml. After homogenization and centrifugation (20,000
g, 10 min), an aliquot of 5 ml was incubated with about 100 g of amyloglucosidase (100 U/mg of enzyme) for 30 min at 60C. After dilution with an aqueous
solution of rhamnose used as internal standard (final concentration of about 510 ppm), 25 ml were injected on the Carbopac PA1 column. This analysis gives
the amount of the free sugars as fructose (Ff), sucrose (S) and glucose as the
sum of free glucose (Gf) and glucose from maltodextrins (Gn). Another aliquot
of 5 ml was treated with Novozym SP 230 (protein/substrate ratio of about
0,01) and amyloglucosidase (100 g) at 60C. This analysis leads to the amount
of total glucose (Gt): Gt Gi Gs Gf Gm, with Gi glucose from inulin
or oligofructose and Gs glucose released from sucrose ( S/1.9) and to the
amount of total fructose (Ft): Ft Fi Fs Ff with Fi fructose released
from inulin or oligofructose and Fs fructose from sucrose (S/1.9).
The inulin or oligofructose content (I) is obtained by: I k (Gi Fi),
with k [180 162 (n-1)]/180 n, n being the sum of fructose and glucose
residues per molecule of inulin or oligofructose. As the general formula of inulin
is GFn and fructose and glucose have the same molecular weight, the number n
may be calculated from the ratio of fructose/glucose amount measured after complete hydrolysis. The value for n of commercial inulin from chicory is around
10. For oligofructose, where the general formulae can be GFm and Fm, a value
for n of about 4 can be safely used.
Results
Tentative determination of oligofructose and inulin in the soluble fiber fraction obtained by the AOAC method: Soluble and insoluble dietary fiber contents
of two foods enriched in oligofructose (filling and yogurt) and of a food enriched
in Raftiline (chocolate bar) were measured by the AOAC method. Table 3
shows that very low values of fibers were obtained for filling and yogurt. The
soluble dietary fiber amount of chocolate bar was higher (9.1 g/100 g wet matter).
The respective amounts of oligofructose and inulin were measured in the soluble
dietary fiber fraction. The recoveries of oligofructose in the case of filling and
yogurt were extremely low. An average degree of polymerization n of 20 was
calculated leading to a value of 0.905 for the factor k used for calculations of
inulin content in the soluble dietary fiber fraction of the chocolate bar. The recovery of inulin, though more important, was less than 32 g/100 g demonstrating
that the conditions used to eliminate starch and proteins and/or to extract the
soluble fiber are too harsh for a quantitative recovery of inulin in the soluble
fraction.
These results show that the AOAC method, in the case of inulin-rich products, seriously underestimates soluble dietary fiber amounts due to the loss of
virtually all oligofructose and an important fraction of inulin.
224
Dysseler et al.
All Bran
Pectibran
26.6
5.3
31.9
30.3
8.5
38.8
28.7
17.2
45.9
28.9
14.2
43.1
(2.5)
(0.8)
(3.1)
(1.1)
(0.5)
(1.6)
(0.5)
(0.3)
(0.8)
(1.3)
(0.6)
(1.7)
238
Craig et al.
Overall, ingestion of polydextrose delivers a number of physiological effects consistent with other DFs. Additional studies are underway.
ANALYTICAL MEASUREMENT OF DF
AOAC Method
The AOAC assay for TDF is a widely accepted protocol in many countries
(10,11). However, this enzyme-gravimetric procedure does not include RO (including polydextrose and inulin) as DF. A schematic of the critical steps in the
AOAC assay is shown in Figure 3. Petroleum ether removes fat, enzymes remove
starch, starch dextrins and protein. The ethanol step is designed to precipitate
polysaccharides, leaving behind sugars and other small molecules. However, the
ethanol precipitation step leaves most RO in the discarded supernatant, along
with sugars. As RO is increasingly used in foods, this step becomes more problematical. In addition, studies (4244) have indicated that a number of polysaccharides (pectin, arabinan and arabinogalactan) are not quantitatively precipitated
by 80% ethanol.
240
Craig et al.
FIGURE 4 HPLC prole of litesse polydextrose (before and after ethanol fractionation).
About 35% precipitated (Fig 8b), leaving 65% in solution (Figure 8b). Fig. 9
compares Raftiline to inulin (from chicory root, Sigma) and to Raftilose (oligofructose). Orafti and INRA have published an HPLC method (45) to specifically
measure -fructans (inulin and oligofructose) in food. They propose that this
value then be incorporated into the DF value.
HPLC Methods
An assay used currently in Japan to quantify polydextrose as fiber utilizes HPLC
(12). Enzymes (-amylase and amyloglucosidase) break down starch/maltodextrins in the sample, similar to the AOAC assay. The sample is then membrane
257
factor to compensate for the difference in degradation during the hydrolysis procedure between free galacturonic acid in the standard solution and polymeric
galacturonic acid in the sample. The use of this factor resulted in better agreement
with other similar methods. It is now included in the published method (9).
The intercomparison study was designed to eliminate as many potential
variables as possible, and some materials were supplied by the coordinator. It
had originally been intended to supply the Uppsala participants with the same
enzymes as the AOAC participants. However, it was found that the amyloglucosidase issued to the AOAC participants contained some unwanted hemicellulase
activity. This caused losses in the Uppsala procedure, which has longer incubation stages. The Uppsala participants were therefore supplied from a purer source.
This illustrates the importance of checking the activity of the enzymes in the
method to be used.
Preliminary Intercomparison Analyses
Before starting the main intercomparison analysis, the participants were asked to
analyze a mixture of four polysaccharide materials arabinogalactan (larch gum),
fibrous cellulose, sugar free citrus pectin, and galactomannan (alcohol washed
guar gum), and to obtain results within the known value 2 SD (based on the
performance in the intercomparison study). Participants who were using gas chromatography were asked, in addition, to carry out a similar exercise on a mixture of
sugars. These two preliminary tests proved to be very useful, as they highlighted
problems in some laboratories with the filtration step of the AOAC procedures,
miscalculations, misunderstandings of the role of blanks, problems with response
factors in GC, a problem with the makeup of a sugar standard and of the unknown
sugar mixture, and uncovered minor errors in the protocol.
Because of this exercise, many laboratories subsequently produced very
good data for the main intercomparison study by all the methods.
Table 2 shows the very low coefficients of variation between laboratories
achieved with all the methods, and the good agreement between the mean values
for dietary fiber. This agreement using the polysaccharide mix contrasts with the
large difference in dietary fiber values found between the Englyst method and
AOAC methods for many food materials and underlines its applicability for initial
quality control purposes.
Main Intercomparison Analyses
The choice of suitable test materials for the study was limited. The five new
reference materials could not be used because the participants must eventually
carry out the analysis for certification without knowing the expected results. Alternate materials had to be found. Four dried reference materials that had already
been tested for homogeneity and stability were kindly supplied by the Food Re-
261
ticipants using the other methods achieved good agreement with the expected
values for the polysaccharide mix. The agreement between methods for the polysaccharide mixture was closer than in the intercomparison studythe mean values range between 91.7 and 93.3 g/100 g d.w. basis.
ResultsEnzyme Testing
The enzymes used by the participants were not supplied by the coordinator, to
avoid systematic bias. The enzymes were tested for activity and purity. Some
participants using the AOAC methods obtained low recoveries from the substrates arabinogalactan, pectin and -glucan when following the enzyme testing
method in the AOAC procedures (985.29 and 991.43). In some cases the laboratory had not dried the substrate or allowed for the substrates moisture content.
This requirement is not clear in the AOAC methods. When allowance was made
for moisture, the recovery was acceptable. However in other cases, recovery was
still low after allowing for moisture, although other participants using the identical batch of enzyme obtained acceptable recoveries. One possible reason could
be a loss of precipitated material through the Celite bed due to the physical nature
of the precipitate and the preparation of the Celite bed. With a small quantity of
pure substrate the risk of losses is probably greater. One participant commented
that it was necessary to allow two hours standing time for the precipitate to develop before filtration to obtain good recoveries, rather than one hour as stated
in the procedure.
ResultsNew Reference Material
Dietary fiber values for the five new reference materials are shown in Tables 5
8. These results are preliminary. They have yet to be amended after discussion
and evaluation at the meeting of participants.
The standard deviation of the mean of means was low, enhancing the possibility of certification. The agreement with the results obtained in the previous
homogeneity study was mostly good, within one standard deviation, with the
exception of apple. In making this comparison, it must be remembered that the
homogeneity analysis was only carried out by one laboratory per method. Agreement between the AOAC Prosky method 985.29 and the AOAC Lee method
991.43 was good for haricot, soya and Bran breakfast cereal, but outside two
standard deviations for carrot and apple (Table 5).
The standard deviation of the mean of means was again low, which is
an improvement on previous studies using the Englyst methods (Table 6). It is
anticipated that it will be possible to certify on the basis of these results. The
agreement with the homogeneity study is very good; all the results are within
one standard deviation. The agreement between the gas chromatographic and
colorimetric methods is good (within 1 SD), except for carrot. This may suggest
that the two methods should be certified separately.
263
AOAC
985.29
(Prosky)
AOAC
991.43
(Lee)
AOAC
991.43
APP**
RM 514
Haricot
RM 515
Carrot
RM 516
Apple
RM 517
FF Soya
RM 518
Bran B/C
23.9 0.8
(25.9)
30.3 0.2
(31.0)
16.2 0.1
(16.4)
12.5 0.1
(12.4)
30.2 0.1
(30.5)
24.9 0.3
(26.1)
29.5 0.2
(29.5)
16.2 0.2
(14.9)
12.8 0.6
(12.6)
29.7 0.2
(30.5)
24.6 0.2
29.4 0.1
16.2 0.3
13.2 0.1
31.0 0.3
Englyst GC
1988
Mongeau
992.16
22.6 0.4
(19.8)
27.2 0.5
(27.1)
15.5 0.6
(13.7)
14.8 0.2
(11.9)
27.3 0.7
(24.3)
20.6 0.8
25.1 0.1
14.6 0.4
14.2 0.2
29.5 0.2
The standard deviation of the means was low. There are some differences
from the values with the AOAC methods, particularly with haricot beans. The
AOAC methods gave approximately 26 g/100 g and the Uppsala methods gave
23.5 g/100 g (Table 7).
Overall, the results show the expected difference between methods based
on the knowledge that they measure different things.
SUPPLEMENTARY STUDY
Additional analyzes were carried out on the new reference materials by Dr.
Mongeau of the Bureau of Nutritional Sciences, Ottawa, using a range of methods
in his laboratory.
The following methods were used. The traditional Prosky method, AOAC
985.29 (2); the AOAC 991.43 method of Lee using MES-TRIS buffer (10) and
a modification using a porcine pancreas enzyme preparation (12); the 1988 version of the Englyst gas chromatographic method (13); and the Mongeau/Brassard
rapid enzymatic-gravimetric method (AOAC 992.16) that includes an initial autoclaving step (14).
The AOAC methods gave very similar values. For carrot, this contrasts
270
Henshall
iodide, cyanide (24). Carbohydrates are easily oxidized on gold or platinum electrodes at high pH and are therefore good candidates for electrochemical detection.
However, DC amperometry is not useful for carbohydrates since the carbohydrate
oxidation products foul the electrode causing a rapidly decreasing response on
subsequent determinations (Figure 2). This problem is solved by pulsed amperometry which utilizes a triple pulse sequence rather than a constant potential on
the working electrode (1-3). Figure 3 illustrates the basic principle of PAD. By
using a repeating sequence of high positive (E2) and negative potentials (E3) following each measurement, a clean electrode surface is maintained which ensures
consistency in response. The detector signal for a particular analyte is measured
at a potential which is appropriate for the particular analyte (E1) by integrating
the current for a fixed length of time (typically 200 ms) and storing the resulting
charge in a sample-and-hold amplifier until the next measurement.
The potential settings for a particular analyte are best determined by using
cyclic voltammetry or pulsed voltammetry (25). Cyclic voltammetry plots such
as the one shown in Figure 4 for glucose on a gold electrode also throw light on
the mechanism of the electrochemical processes in pulsed amperometry, and the
reason for the high specificity of the technique. The solid line shows the variation
in current as the potential is swept from 0.8V to 0.6V and back to 0.8V for
glucose in 100 mM NaOH. The dashed line shows the current as a function of
FIGURE 2 Decrease in detector response for repeat injections of a carbohydrate and an amino acid (concentration 10 g/L in both cases) using DC amperometric detection. Peaks: (1) leucine. (2) lactose. Column: CarboPac PA1 (10
* M, 250 4 mm i.d.) with guard, Eluent: gradient, 15 to 150 mM NaOH in 5
min. Detection: DC amperometry, Au electrode, range 1*A, applied potential
0.15V vs. Ag/AgCL.
HPAE-PAD
273
PKa
Fructose
Mannose
Xyolse
Glucose
Galactose
Dulcitol
Sorbitol
-Methyl glucoside
12.03
12.08
12.15
12.28
12.39
13.43
13.60
13.71
276
Henshall
cles. The extremely small diameter of the pellicular beads results in excellent
mass transfer characteristics and highly efficient separations. This structure also
permits easy scale up from analytical columns (4 mm) to the semi-preparative
scale CarboPac columns (9 mm, and 22 mm).
HPAE-PAD
277
FIGURE 10 Separation of maltose oligomers by HPAE-PAD. Peak labels indicate degree of polymerization (DP). Column: CarboPac PA100 (8.5 M, 250
4 mm i.d.) with guard. Eluent: NaOH/sodium acetate gradient. Flowrate:
1.0 mL/min. Detection: pulsed amperometry, Au electrode.
FIGURE 12 Separation of a hydrolyzed corn syrup by HPAE-PAD using gradient elution. Column: CarboPac PA1 (10 M, 250 4 mm i.d.) with guard.
Eluent: 150 mM sodium hydroxide with sodium acetate gradient. Flowrate:
1.0 mL min. Temp: ambient. Detector: pulsed amperometric, Au electrode.
Peaks: 1 glucose, 2 DP 2, 3 DP 3, 4 DP 4, 5 DP 5.
HPAE-PAD
279
FIGURE 13 Chain length distribution of debranched amylopections from various sources determined by HPAE-PAD. column: CarboPac PA1 (10 M, 250
4 mm i.d.) with guard. Eluent: A, 150 mM NaOH; B, 150 mM NaOH/500 mM
sodium acetate. Gradient program: 40% B at 0 min, 50% at 2 min, 60% at 10
min, 80% at 40 min. Temp: ambient. Flowrate: 1.0 mL/min. Detector: pulsed
amperometric, Au electrode, range 10 A. Adapted with permission from
Koizumi, K., Fukuda, M., Hizukuri, S., (1991), J. chrom. 585, 233238.
QUANTITATION OF POLYSACCHARIDES
For exact quantitation of each polysaccharide, the variation in PAD response
over the DP range of interest is required. Relative detector response for maltose
oligomers in the range DP 2 to DP 7 determined by Koizumi et al. are shown
in Table 2 (20). For the maltooligomers in the range of DP 6-17, the relative
282
Henshall
FIGURE 15 determination of ketose and related sugars by HPAE-PAD. Column: CarboPac PA1 (10 M. 250 4 mm i.d.) with guard. Eluent 100 mM
NaOH/20 mM sodium acetate. Flowrate: 1.0 mL/min. Temp; ambient. Detector: pulsed amperometric, Au electrode, range 1 A. Peaks: 1 galactinose,
2 sucrose, 3 1-ketose, 4 6-ketose, 5 neo-ketose, 6 nystose.
the activity of putrefactive bacteria and reduces the formation of toxic fermentation products. In Japan, oligosaccharide additives are used in over 450 food products (27) primarily for their beneficial health effects. These products are typically
mixtures of small oligosaccharides such as the fructooligosaccharides shown in
Figure 14. Analysis of these types of products is straightforward using HPAEPAD as shown in Figure 15. Oligofructose products derived from chicory inulin
having DP values up to 60 can also be determined by HPAE-PAD using gradient
elution as shown in Figure 16. Figure 17 shows the separation of Raftiline, a
commercial inulin product from a European manufacturer in which the smaller
oligofructose (Fn) chains are distinguished from the inulin (GFn) chains. Response factors have also been determined for the Fn and GFn oligomers up to
DP 8. Determination of response factors for higher DP oligomers is in progress
(H. Hoebregs, private communication).
284
Henshall
with a run time of less than 13 min. Figure 18 shows results obtained for a citrus
pectin hydrolysate and a beet hydrolysate (11).
It is also possible to determine neutral sugars and uronic acids in a single
run by using gradient elution (J. Prodolliet, Nestle, Lausanne, personal communication). The determination of neutral sugars and uronic acid constituents by
HPAE-PAD is now accepted as an alternative method for nutritional labeling
purposes in the UK (H. Englyst, personal communication).
288
Henshall
ACKNOWLEDGMENTS
The author wishes to acknowlege the contribution of the applications and research
staff of Dionex Corporation who carried out the chromatographic separations
shown in Figures 2, 912, 15,16, and 20.
REFERENCES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
296
Kays et al.
and snack foods (24, 25); beta-glucan in barley (26); starch in grains (27), pulses
(28), and snack foods (25); starch damage in flour (29); amylose in rice (30);
sucrose in chocolate (31) and wine (32); soluble solids in vegetables (33); and
alcohol in beer (34) and wine (32).
Product
Breakfast cereals (24)
Snack food (25)
Oat bran products (22)
Calibration
Prediction
Fiber
fraction
R2
SE
SE
NDF
NDF
IDF
SDF
TDF
TDF
40
40
45
45
45
90
0.98
0.82
0.99
1.5
1.8
1.6
50
49
24
24
24
29
0.96
0.83
0.92
0.92
0.97
0.99
1.8
1.4
1.0
0.34
0.85
1.5
NDF, neutral detergent ber; IDF, insoluble dietary ber; SDF, soluble dietary ber;
TDF, total dietary ber
b
n, number of samples; R 2 and 2, coefcients of determination for calibration and prediction, respectively; SE, standard error
NIR Analysis
297
foods such as potato chips, pretzels, and popcorn (25). A two term, first derivative
equation using 4 wavelengths was developed relating NDF to NIR reflectance
spectra. The wavelengths chosen, via stepwise multiple regression analysis, were
2364, 2088, 1936, and 2342 nm and are indicative of fiber, starch, water and fat
and/or protein absorbance, respectively. The poorer correlation (Table 1) was
partly attributed to a skewed distribution of NDF values (62% of samples had
NDF values below 2%) and partly to the inclusion of cornmeal products which,
as a group, did not predict well. The authors suggested that cornmeal based products may require a separate calibration. If cornmeal samples were omitted, the
standard errors for calibration and prediction are reduced to 1.4% and 1.3%, and
the correlations improved to 0.91 and 0.89, respectively. Collectively the data
indicate a potential relationship between insoluble fiber, as measured using the
NDF method, and NIR spectra in snack foods and reiterate the importance of
sample selection and a relatively normal distribution of sample values.
Williams et al. assessed the potential of analyzing total, soluble, and insoluble dietary fiber in oat bran products using NIR reflectance spectroscopy (22).
Oat bran is an excellent source of soluble dietary fiber which has been associated
with several health benefits. As a consequence there is interest, among manufacturers of oat based foods, in the rapid measurement of components of dietary
fiber for composition assurance. Total dietary fiber and its constituents, soluble
298
Kays et al.
and insoluble fiber, were determined in the laboratory by the method of Mongeau
and Brassard (4). Using an NIRSystems scanning monochromator for the collection of spectral data and NSAS NIR software, models were developed to predict
soluble, insoluble, and total dietary fiber content of oat bran products. The accuracy of NIR prediction of the components of oat bran fiber, using PLS regression
analysis, was deemed adequate for quality assurance (Table 1). A comparison
between stepwise/multiple linear regression, in which specific wavelengths were
selected, and PLS regression, which uses the entire NIR spectrum, indicated that
the two regression methods gave very similar calibrations for soluble and insoluble dietary fiber (22). However, in the case of total dietary fiber PLS regression
was the better method, having a higher 2 value and lower standard error of
performance during verification.
Using wheat bran products only, Horvath et al. investigated relationships
between NIR spectra and dietary fiber constituents (39). An enzymatic method
(40) was used for water soluble and insoluble dietary fiber determination and
constituents such as lignin, cellulose, hemicelluloses, and pectin were determined
chemically within the dietary fiber fractions. Linear regression of a second derivative transformation of log10 1/R spectra was used to identify optimum wavelengths for predicting the soluble and insoluble dietary fiber and total dietary
fiber. Correlation coefficients obtained for all constituents in this narrow population were 0.95 or greater with standard errors of calibration that were adequate
for the state of the art at that time. Validation statistics were not reported.
The cereal and baking industries market products with a wide range of
cereals often with multiple grains in a single product. Furthermore, regulatory
agencies obtain samples from many sources, therefore, any method used in monitoring the products must be capable of assessing a broad range of samples and
values. Kays et al. developed a calibration for predicting total dietary fiber in a
variety of cereal products with a broad range of dietary fiber values (23). Both
processed and unprocessed wheat, oats, rye, barley, amaranth, corn, and rice were
included with an overall range in total dietary fiber values from 1 to 52% (n
90). The samples contained 10% fat and/or 20% sugar. Higher fat/sugar
samples were avoided due to difficulty in grinding and interference with the reference method. The reference method used to measure total dietary fiber was the
AOAC procedure 991.43 (41), recommended for food labeling in the United
States. The method includes non-starch polysaccharides, lignin, resistant starch,
cutins, and plant waxes in the dietary fiber measurement. Spectral reflectance data
were collected from 90 ground samples using an NIRSystems scanning monochromator and Infrasoft International software. The spectral data were transformed with standard normal variate and detrending to remove multiplicative
interferences of scatter (42), and then transformed with second derivative processing. PLS was the regression method selected to relate spectral data to laboratory reference values and cross-validation was used to determine the optimal
NIR Analysis
299
number of PLS factors for dietary fiber prediction. A NIR calibration equation
was obtained for the prediction of total dietary fiber and used to determine the
total dietary fiber content of samples in an independent set (Table 1, Figure 2).
Linear regression of AOAC determined dietary fiber against NIRS predicted dietary fiber gave an intercept and slope not significantly different from 0.0 and
1.0, respectively.
The number of factors (nine) in the calibration developed was partly a
function of the diversity of the data set. The calibration developed has broad
utility for manufacturers using multiple grains and multiple grain products and for
regulatory agencies monitoring products from many sources. Many companies,
however, market products that contain only one or two types of grains and for
these industries a simpler model may suffice. When samples containing oat and
wheat only were utilized, a new calibration was obtained (n 42) with only
four factors and similar standard error of cross validation, coefficient of determination and accuracy of prediction as the model derived from the broader data set
(23). Thus, calibration development and sample prediction were achieved for
determination of total dietary fiber in a heterogeneous group of products and in
a more homogeneous group.
Loadings express the spectral variation that corresponds to each factor of
the PLS model and can provide information on the constituents exerting the greatest influence for each factor. Intensity of absorption peaks in the loading spectra
FIGURE 2 AOAC determined total dietary ber versus NIRS predicted total
dietary ber for cereal products in the calibration (A) and validation (B) data
sets (adapted from Ref. 23)
329
Insoluble Fiber
The most common form of dietary fiber is insoluble fiber which is composed of
cellulose, lignin, and some hemicellulose. It promotes regularity and is being
studied for its potential for reduce risk of colon/rectal cancers. Although cereal
brans and whole-grain cereals are the most commonly recognized source, other
good sources of insoluble fiber are dried beans, peas, vegetables, and nuts.
Soluble Fiber
Soluble fiber comprises about 25% of the dietary fiber consumed but strong evidence shows that soluble fiber as part of a low-fat, low-cholesterol diet may help
lower blood cholesterol in those individuals with elevated blood cholesterol levels. It has also been shown to help control blood glucose levels in individuals
with diabetes mellitus. Good sources of soluble fiber are whole-grain oats and
barley, oat bran, some fruits, dried beans, and other legumes. According to current
nutrition labeling regulations dietary fibercontaining food must have at least 2.5
grams fiber per reference serving to make a good source claims and at least
5.0 grams of fiber to make an excellent source claim.
340
Dreher
Mechanical/Physical
Microstructural
Functional
Examples
Cohesiveness
Hardness
Brittleness
Gumminess
Density
Expansion
Shear
Viscosity
Cellularity
Crystallinity
Porosity
Uniformity
Water binding
Emulsification
Spreadability
Whippability
Industry Availability
A wide variety of dietary fiber sources are available to the food industry. Since
the early 1980s, the number and availability of dietary fiber sources has increased
360
Dreher
prior to use. Other ingredients are also useful in improving loaf volume. The
addition of shortening and/or surfactants (e.g., sodium stearoyl lactylate) helps
to increase the loaf volume of bran supplemented breads.
Corn Bran
There are two types of corn milling processes (Hoseney, 1986). Dry milling primarily separates the anatomical parts of the corn: bran, germ, and endosperm.
Wet milling separates corn into its chemical constituents: starch, protein, oil, and
fiber. Although most corn is processed by wet milling, dry milling is typically
used for the production of corn bran for human consumption (Burge and Duensing, 1989). Because of density differences between germ and bran, raw corn
bran can be aspirated from the bran/germ stream for further refining, concentrating, and sizing. This results in a very high fiber corn bran product with about 85
percent or more dietary fiber which is about 65 percent hemicellulose and 20
percent cellulose. Its calorie content is low at about 0.5 kcal/g. Corn bran exhibits
a water binding capacity of about 2.4 to 3.7 g/g depending on the particle size
and dietary fiber content. This product comes in various particle sizes, from fine
to coarse, as needed for a variety of specific food applications.
Rice Bran
Rice is in the form of paddy (where the kernel is fully enveloped by the rice
hull) when harvested from the field (Saunders, 1990). After being dried, the first
stage in milling is removal of the hull, yielding brown rice. In the second state,
the outer brown layer is removed from the brown rice kernel by abrasive milling
operation to yield the familiar white rice. The separated brown layer is designated
rice bran, which includes the germ in the United States. Under normal milling
conditions, when brown rice is milled to white rice, the oil in the bran and a
potent lipase also in the bran come into contact, resulting in rapid degradation
of the oil. The bran thus produced is unpalatable and only fit as a feedstuff.
However, if the bran is subjected to a short-term high temperature heat treatment
the lipase activity is destroyed and a stabilized bran is produced. The compositions of rice brans are summarized in Table 5.
Rice bran has a number of unique properties (Saunders, 1990). Rice bran
protein is of relatively high nutritional value. Protein efficiency ratio values reported for bran generally range from 1.6 to 1.9 compared to casein value of 2.5.
Digestibility of protein in rice bran is about 75 percent. The bran usually contains
1622 percent oil, although this value is higher in parboiled bran due to the
absence of broken starch fragments. The three major fatty acids in rice bran are
palmitic, oleic, and linoleic and they make up more than 90 percent of the total
fatty acids. Rice bran oils contain 34 percent waxes and about 4 percent unsaponifiable lipids. Rice bran oil has a higher content of unsaponifiable material
375
to modify the fiber molecules in each co-product (bran and flour). Table 1 shows
the dietary fiber composition of common wheat products. All-purpose and cake
flours have substantial amounts of fiber removed during milling.
Oat gum extracted from oat bran with aqueous sodium carbonate (pH 10)
at 40C had different properties depending upon the concentration procedure used
(7). Dialysis of the extract produced gums with greater viscosity than did ultrafiltration or ethanol precipitation methods. Alcohol precipitation had the highest
yields, however. Inactivation of enzymes by 75% ethanol at 80C also resulted
in gums of higher viscosity but much of the -glucanase present in the original
bran had been reduced by a prior steaming by the supplier.
Alkali extractions were also effective in removing soluble fiber from defatted rice bran. Extracts obtained with either sodium hydroxide or calcium hydroxide at pH 12 significantly lowered serum cholesterol in rats fed a cholesterolenriched diet (8). The Ca(OH) 2 method was preferred because it produced a
lighter colored-product. NaOH had a better yield, however, of 8 g per 100 g rice
bran versus 5 g for the calcium salt.
Removal of starch, protein and other water-soluble compounds by various
treatments affected wheat bran composition, water-holding capacity, and breadbaking properties (9) (Figure 1). Amylase treatments produced the highest neutral
fiber contents; protease with amylase yielded more fiber, but that fiber was not
suitable for baking. Rinsing following chemical or enzyme treatment may have
removed too much soluble material.
Microwave heating has been proposed as a pretreatment for fruit waste to
enhance pectin extraction. A ten-minute treatment at 2450 MHz and 0.5 kW
improved pectin yields from orange and lemon peels and apple pressings without
affecting the amount of anhydrouronic acid content or degree of esterification
(10). Microwave heating, whether or not followed by drying, increased pectin
gel strength compared to pectin extracted directly from fresh fruit or from dried
TDF (%) a
12.2
2.7
1.7
42.8
13.2
381
SUMMARY
Common processing methods can produce important changes in food dietary fiber
composition. These changes are summarized in Table 2. Although calculation of
nutrient content of processed foods from ingredient content is an accepted procedure in the United States, dietary fiber composition will most likely be incorrect.
New sterilization methods such as ohmic heating and magnetic field processing
may also affect fiber. As new methods are developed, care should be taken to
analyze the fractions of fiber, not simply total fiber. Processing-induced changes
in fiber molecules may significantly alter health benefits, leading to public health
and food labeling issues.
388
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389
Several patents have been filed over the last six years for the production
of RS for use in food as bulking agent, dietary fiber, or fat mimetic (3440).
Generally starch is heated in water at more than 100C to hydrate and gelatinize
starch and then cooled for a sufficient time to cause starch retrogradation and
formation of RS fractions. The yield of RS depends on the amylose content, and
consequently the preferred sources are starches having 70% or more amylose
content (amylomaize with 70 or 85% amylose, high amylose pea starch and high
amylose barley starch). Starch is mixed in 3 to 10 volumes of water, but the
highest RS yield is achieved with greater starch concentration (4,3437). The
suspension is heated in an autoclave at a temperature above 100C to ensure
starch gelatinization and complete disruption of the granules. High temperatures
are particularly necessary for the high amylose starches (120150C). In one of
the proposed processes (4,34), the mixture is then cooled to below room temperature to allow amylose retro-gradation to take place and the heating-cooling
process is repeated up to three times (Table 2). The yield of RS is systematically much lower with method B used for RS determination compared with the
milder pancreatic -amylase assay shown in the last column. Storage at 4C for
periods of 224 hours is recommended to complete amylose reordering. RS
contents increased when heating from 121 to 134C, and decreased again at
148C (4). The yield of RS shown in Figure 1 is inversely related to the amylose content.
RS3 has been produced by extrusion cooking of amylomaize starch in a
double screw extruder with one volume of water added, followed by two days
storage at 4C, oven-dried and milled. The in vitro RS content was 30% (Table
3), but reached 49% in ileostomy subjects (19). This partially retrograded starch
was used as the reference sample in the Euresta project. By autoclaving at 145C
392
Wursch
readily available source of RS, or dietary fiber. Breads and breakfast cereals enriched with Hi-maize are already on the market in Australia (39,40). Bread has
been enriched up to 11% RS (based on DM, method A) by replacing part of the
flour by retrograded amylomaize (70% amylose) or native amylomaize and gluten
without altering the texture and taste of the product (45). These application studies show that RS content in amylomaize is markedly lower, around 3040%,
after the processing of the food (Table 3).
Chemical modification of starch (CMS) can be another way to significantly
reduce its digestibility. Chemically modified starches are widely used in the food
industry as texture agent. Chemical modification such as substitution and crosslinking confers to the starch heat and shear resistance, and/or improved pH and
freeze-thaw stability. The in vitro digestibility of acetylated and cross-linked gelatinized starches was nearly equal to native gelatinized starches (47), whereas
etherification with hydroxypropyl groups significantly reduced the digestibility
(47, 48). Increasing molar substitution (MS) decreased the in vitro and in vivo
digestibility to around 42% for a MS of 0.16 (49). The undigested fraction was
metabolized in rats only to a very limited extent due to the very high MS of this
residue (DS 0.6) (49). These studies and others indicate that only substitution
with ether groups decreases the digestibility of gelatinized starch, but the resulting
resistant starch is not metabolized by the intestinal microflora.
CONCLUSIONS
Resistant starch has been known for 15 years, and its chemical and physical
natures have been well defined. The increase in RS in everyday food by processing adaptation is quite limited, thus the attention has been concentrated on
high-amylose starch, particularly from corn, which is itself in great part resistant
to digestion. Starches with 50% RS or more when the starch was purified can
be produced, and they withstand heat treatment. These starches can be easily
incorporated in food to increase the fermentable polysaccharide content without
altering the sensory properties. RS is slowly fermented in the large intestine by
the colonic flora. The metabolites formed include volatile fatty acids (acetic, propionic and butyric acids), resulting in a decrease in the pH of the feces (49,50).
RS seems to yield more butyrate than some highly fermentable soluble fibers,
which is of special interest regarding the prevention of colonic cancer (51).
REFERENCES
1.
2.
3.
4.
Englyst, H., Wiggins, H.S. & Cummings, J.H. (1982) Analyst 107, 307318.
Siljestrom, M. & Asp, N.G. (1985) Z. Lebensm. Unters forsch. 148, 48.
Berry, C.S.J. (1986) Cereal Sci. 4, 301314.
Sievert, D. & Pomeranz, Y. (1989) Cereal Chem. 66, 342347.
398
Rabe
with 2 M KOH or DMSO. RS was the starch solubilized in the DF residue or the
difference in starch content with and without added DMSO. Different methods for
starch determination were used, too. In products with a high starch to fiber ratio,
formation of enzyme-resistant starch during processing may contribute significantly to the DF content of the product. RS has been identified in cooked and
baked foods such as bread, boiled potatoes, pasta, legumes, porridge, and corn
flakes at levels of 14 g/100 g d.m. Cooked, dried, and ground legume seeds,
such as beans, which are rich in amylose, showed an RS content of 36%, which
is considerably higher than that formed during baking of bread. As the raw beans
showed 0.8% RS it may be that these products have a high content of RS2 (12,13),
which is resistant starch like raw potato starch.
Although Englyst (14) proposed a method for the determination of different
kinds of RS, which was accepted initially, there is still the need for a quick and
reliable method, mainly for the determination of RS3, the retrograded amylose.
The true RS content of foods may be higher than that recorded for RS3 in fiber
residues since only a fraction of retrograded starch remains in the residues. The
continuing development of new methods (15) shows there still is demand for it.
Although interesting from a scientific point of view, the physically inaccessible
kinds of RS are not of great importance, as most food is consumed after heating.
To take this to the extreme, it means RS2 from potatoes should not be considered,
because when they are consumed, there is only RS3. On the other hand, in eating
raw cereals like muesli, there really is a greater consumption of RS2, physically
trapped starch, but only a small part of the population consumes it.
The repeated heating of starch-rich foods (Table 2), measured by the IDF
fraction, shows the possibility of accumulation of RS. Potatoes (variety
Hansa) were cooked in water, drained, cooled to room temperature, and stored
in a refrigerator for 18 h (overnight). Heating of the cooked potatoes, the cooked
polished rice, and the pasta (100% durum wheat) in the microwave oven was
done without additional water.
It can be seen from Table 2 that the content of IDF, or of RS, respectively,
is greater in products heated with excess water in a normal cooking pot than in
products heated without water, e.g. in a microwave oven, as expected. On the
other hand, in heating in the microwave oven, there was an increase, too. The
highest amount of RS, compared to the original but low value for the uncooked
food, was formed in rice heated in a normal cooking pot or a microwave oven.
In pasta, too, there was a great increase, but only about half that in rice. The
reason was that these foods are rich in amylose and underwent complete starch
gelatinization during cooking. The least amount was formed in potatoes, independent of the method of heating. The decrease in RS in rice during heating in the
normal pot results from the fact that in this case, there was a great amount of
insoluble material poured away with the cooking water, which was opaque from
it (16).
415
Polydextrose
Polydextrose is prepared by vacuum thermal polymerization of glucose, using
sorbitol as a plasticizer and citric acid as a catalyst (25). Although 16 glycosidic
bonds predominate (both alpha and beta), polydextrose is composed almost entirely of randomly cross-linked glucose polymers. The energy value of polydextrose is only 1 kcal/g because the random bonding renders it relatively resistant
to digestive enzymes, with a large proportion excreted unchanged in the feces
(26). Because of the low digestibility of polydextrose, laxative effects have been
observed above the threshold value of 90 g/d (26). No deleterious effects of
polydextrose have been observed in animal testing, and it does not appear to
interfere with the absorption and utilization of essential vitamins, minerals, or
amino acids (27).
Polydextrose exists in five forms, all of which are highly stable (28). The
original polydextrose powder is available in coarse (Pfizer, Inc.) and fine (A. E.
Staley) granulations and has a tart/bitter aftertaste (pH of 2.5 to 3.5). Type N
polydextrose is a 70% solution neutralized with potassium hydroxide to a pH of
5.0 to 6.0 and has been described as being slightly sweet-salty-bitter (2628).
Type K polydextrose is a powder buffered to the same pH as type N with potassium bicarbonate. In solution, polydextrose type K has a slightly sweet-sourbitter taste (30). The fourth form, type F (Litesse, Pfizer, Inc.), is an improved
polydextrose powder with reduced titratable acidity and a bland/neutral taste.
The recently developed Litesse II (Pfizer, Inc.) has tenfold lower titratable acidity than Litesse and has been described as having a clean, mildly sweet taste.
With lower acidity levels, Litesse II can be used at much higher concentrations
than previous versions of polydextrose and has negligible sucrose inversion and
minimal effects on fat rancidity (26).
As a low-calorie bulking agent, polydextrose reduces the calories of fullsugar foods while maintaining body and texture. Polydextrose can also contribute
to the mouthfeel and creaminess of fat-reduced formulations because of its high
viscosity in solution (26). Because polydextrose slows moisture changes in foods,
its use in dry mixes improves flowability, assures uniform dispersion, and prevents lumping (28). The essentially bland taste of polydextrose renders it applicable to both sweet and savory applications. However, certain sweet products, such
as confectionary, usually require the addition of a high intensity sweetener like
aspartame, acesulfame-K, or alitame (26). An example of a low-fat confectionary
product containing polydextrose is Lo, launched in 1991 by the Leaf Group in
Finland. Thus, polydextrose has been used in the preparation of bakery goods
417
Maltodextrins
The success of starchy products in reducing the fat content in food formulations
is partially dependent upon having particle sizes small enough to simulate fat
emulsions. Small particle native starches from certain grass and cereal seeds exist,
but their use is uneconomical due to the production costs involved in their preparation (17). For this reason, enzymatic degradation of starch polysaccharides has
been employed to produce maltodextrins as fat replacers. Typical starch sources
are corn, potato, tapioca, and rice. With dextrose equivalents (DE, an indication
of the extent of starch hydrolysis) below 20, maltodextrins are highly digestible
(supplying 4 kcal/g), are easy to blend with other dry ingredients, are readily
dissolved, and have low viscosity in solution (30). Low-DE maltodextrin gels
(consisting of 1 part maltodextrin and 3 parts water) only have 1 kcal/g.
The most useful characteristic of maltodextrins in fat replacement is their
ability to form thermoreversible gels that melt upon heating and reset to comparable gel strength when cooled (31). The main function of the soft and spreadable
maltodextrin gels is to provide texture and mouthfeel simulation of fats in food
products. Maltodextrins may also act as bulking agents. However, like native
starches, maltodextrins have the tendency to retrograde slowly in food products
during storage (31). Low-DE (i.e. DE 5) maltodextrins can also exhibit variable
freeze-thaw stability, unreliable heat and acid stability, and an undesirable starchy
flavor to delicate foods if used at high concentrations (38). Many stable food
starches used as fat replacers are chemically modified to be resistant to temperature, shear, and low pH levels encountered in processing (17,28) (Table 1).
Low-DE maltodextrins derived from potato starch have advantages over
those produced from other starch sources. Potato starch is thought to retrograde
less readily than other starches because of its longer chain amylose molecules,
reducing the tendency for turbidity and undesirable texture (30). In addition to
being thermoreversible, the potato-based maltodextrin gels are pH stable (pH 3
7), can withstand shearing and heating effects, and have a unique plastic,
spreadable, shorthening-like texture (30). Commercial maltodextrins derived
from potato starch include C*Pur 01906 (Cerestar), Paselli SA2 (Avebe), and
Lycadex 100 (Roquette).
Maltodextrins can be used in a variety of products. The shortening-like
texture of potato maltodextrins makes them ideal as fat replacers in baked prod-
423
Slimgel 100/i also has higher stability to a variety of food processing conditions
(56). Slimgel products are used in meat products (Slimgel 100), spreads (usually Slimgel 200), dairy products and desserts, dressings and sauces, and baked
products (56). Slimgel 100/i is preferably used in frozen desserts, dressings,
and sauces because of its cold solubility, ease of use, and improved mouthfeel
due to slower melting (56).
Blends of galactomannans and xanthan are often used in food applications
because they have been found to have functionalities that were at least the same
or better than either gum alone at a reduced cost. When locust bean gum and
xanthan gum are heated together, the resulting solution has a much higher viscosity than would be obtained with either gum alone. At high enough concentrations,
locust bean/xanthan gum blends form thermoreversible gels with a melting temperature between 50 and 55C (57). Guar gum/xanthan gum blends have a synergistic increase in viscosity but without gel formation (58). Kel-Lite CM (Kelco)
is a xanthan/guar gum blend used to eliminate shortening in baked goods through
its stabilizing and emulsifying properties (24).
Galactomannans are also used with certain types of carrageenans. Guar
gum with low concentrations of -carageenan produce an increase in viscosity
whereas high carageenan concentrations increase gel strength. Locust bean gum
and -carageenan enhances gel strength and a range of textures from brittle to
elastic. Locust bean gum/carageenan also lessens the degree of syneresis (23).
FIBERS
A number of fat replacers have been based on fiber from a variety of sources
such as soy beans, oats, sugar beets, almonds, peas, and wheat (Table 1). In
addition to replacing fat, products containing -glucan and inulin have been reported to reduce serum cholesterol (59,60). The fat replacer products Oatrim,
Z-trim, Vitacel, and inulin are described in more detail below.
Oatrim
In 1991, the U.S. Department of Agriculture (USDA) developed an oat-based fat
substitute called Oatrim. (-Amylase enzymes were used to break down the amylose and amylopectin chains in the oat flour or bran to maltodextrins. The enzyme
treatment conditions were selected so that the -glucan bound to the cellular
matrix was liberated, ranging in levels from 110%. The blend of -glucan and
low-DE maltodextrin produced a product with improved functional properties
and fat-like mouthfeel in comparison with other maltodextrin gels. An added
benefit of the -glucan is its reported lowering effects on blood cholesterol and
triglycerides (50). ConAgra Specialty Grain Products and Rhone-Poulenc/Quaker
Oats Companies are currently producing Oatrim under a licensing agreement with